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A targeted siRNA based screen to identify calcium transporters involved in the calcium-dependent regulation of ABCC3 gene expression in a model of breast cancer epithelial-mesenchymal transition (EMT) [Poster]

Citation

Stewart, TA and Azimi, I and Davis, FM and Thompson, EW and Roberts-Thomson, SJ and Monteith, GR, A targeted siRNA based screen to identify calcium transporters involved in the calcium-dependent regulation of ABCC3 gene expression in a model of breast cancer epithelial-mesenchymal transition (EMT) [Poster], Australasian Society of Clinical and Experimental Pharmacologists and Toxicologists Annual Scientific Meeting (ASCEPT2013), 1-4 December 2013, Melbourne, Australia, pp. 132. (2013) [Conference Extract]


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Abstract

Introduction: Invasion and metastasis are hallmarks of cancer and are linked to resistance to anticancer therapies. Increased expression of members of the ATP-binding cassette (ABC) transporter superfamily is also implicated in resistance to chemotherapeutics, and has been linked to invasive behaviour. We have previously identified the Ca2+- dependent upregulation of ABCC3 gene expression in a model of breast cancer epithelial-mesenchymal transition (EMT), a process involved in metastasis. The Ca2+ transporter(s) involved in the regulation of ABCC3 gene expression in this model of EMT are yet to be identified.

Aims: To identify the Ca2+ channels and/or pumps involved in the Ca2+-dependent upregulation of ABCC3 transporter gene expression in a breast cancer model of epidermal growth factor (EGF)-induced EMT.

Methods: To assess the role of Ca2+ transporters in ABCC3 gene expression, an siRNA-based screen targeting 24 Ca2+ channels and pumps using Dharmacon On-TARGETplus SMARTpool siRNAs was performed. MDA-MB-468 basal-like breast cancer cells were transfected with each siRNA SMARTpool for 48 h. Following transfection, cells were serum starved (0.5% FBS) for 24 h before induction of EMT via treatment with EGF (50 ng/mL) for 48 h. Quantitative RT-PCR was used to assess changes in ABCC3 mRNA expression following siRNA and EGF treatment.

Results: siRNA mediated silencing of the Ca2+ permeable ion channel, TRPM7, a previously characterised partial regulator of EMT in this model, did not inhibit ABCC3 gene expression following 24 h EGF treatment. However, siRNA screening identified other Ca2+ transporters that may act as potential regulators of EGF-induced ABCC3 expression at 48 h, for further characterisation.

Discussion: Induction of EMT by EGF in MDA-MB-468 breast cancer cells results in the Ca2+-dependent upregulation of ABCC3 transporter gene expression. Further studies are required to characterise the Ca2+ transporters involved in the regulation of ABCC3 expression in this model.

Item Details

Item Type:Conference Extract
Keywords:EMT, calcium, breast cancer
Research Division:Biomedical and Clinical Sciences
Research Group:Pharmacology and pharmaceutical sciences
Research Field:Basic pharmacology
Objective Division:Expanding Knowledge
Objective Group:Expanding knowledge
Objective Field:Expanding knowledge in the health sciences
UTAS Author:Azimi, I (Dr Iman Azimi)
ID Code:151210
Year Published:2013
Deposited By:Pharmacy
Deposited On:2022-07-25
Last Modified:2022-07-28
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